Recognition Sequence :
GCANNNN↑NTGC
CGTN↓NNNNACG
Unit Definition :
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°C in a total reaction volume of 50 μl.
Reaction Temperature :
60°C
Storage Buffer :
10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; and 50% glycerol
Ligation :
After 5-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Source :
Bacillus stearothermophilus AP
Assayed on :
Lambda DNA (HindIII-digest)
Working buffer :
W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Non-specific hydrolisis :
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 5 u.a. of enzyme for 16 hours at 60°C.
Concentration, u.a./ml :
5000
Inactivation :
20min Under 80°C
Reagents Supplied :
10 X SE-buffer W
Notes :
High enzyme concentration may result in star activity. At 37 centigrade activity is 50% from maximum.
The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,5.
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